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In 2017, WHO published an updated classification of the pituitary adenomas according to the lineages defined by the transcription factors, PIT1, SF1 and TPIT. Nomenclature of the pituitary tumors follows the mature cell types such as somatotroph (GH), lactotroph (LH), thyrotroph, corticotroph, and gonadotroph (FSH, LH). Null cell adenomas are defined by the absence of expression of any hormones and transcription factors. Not infrequently, the pituitary adenomas are invasive to the adjacent structures and are designated as aggressive adenomas. Knosp grading is often used to define the aggressiveness of the tumor. Sparsely granulated somatotroph adenomas and Crooke cell corticotroph adenomas are representative aggressive adenomas. Recently, genomics regarding various adenomas have been clarified, such as GNAS for somatotrophs and USP8 for corticotrophs. Familial pituitary adenomas are another aspect which has been clarified such as MEN1, Carney's complex, familial isolated pituitary adenoma and McCune-Albright syndrome. The pituitary adenomas often produce GH or PRL, hormones of PIT1 transcription factor. It has been agreed that the pituitary adenomas share the characteristics of neuroendocrine neoplasms. The terminology of pituitary neuroendocrine tumor has been discussed. This review article covers various aspects of pituitary adenomas.

Changes in human life style have increased the incidence of obesity which has become a risk factor to human health. In Taiwan, the annual production of Citrus grandis [L.] reaches 75 000 tons, while the amount of waste fruits could reach 5%. We propose that abundant phytochemicals present in the wastes can be beneficial to attenuate obesity with hyperlipidemia. An animal Wistar rat model showed that pomelo flesh, peels, carpel (the segmental membrane), as well as the essential oil attenuated obesity as well as hyperlipidemia, and the AMPK-SREBP-PPARS pathway was involved. To further verify the mechanism, a HepG2 cell model was used to test some representative pomelo phytonutrients including limonene (Ln), γ-terpinene (γT), p-synephrine (SP), β-sitosterol (βS), and hesperidin (Hn). Data interestingly revealed it to be a multiple mechanism of anti-lipogenic bioactivity via downregulating the enzymes involved in both the cholesterol and triacylglyceride de novo biosynthesis (in the order of decreasing bioactivity): acetyl CoA carboxylase (Hn, SP, βS, Ln = γT), fatty acid synthase (Ln, γT, SP, βS, Hn), HMG-CoA synthase (Ln, Hn, SP, βS, γT), and HMG-CoA reductase (Hn, Ln, βS, γT, SP), and via upregulation of cholesterol-7α-monooxygenase (CYP7A1) (Hn, βS). In addition, all pomelo fruit parts enhanced the fecal crude lipid and sterol excretion capability. Thus, fruits of C. grandis can serve as a rather promising integrative antihyperlipidemic agent.

Changing food preferences brought about by westernization that have deleterious health effects1,2—combined with myriad forces that are contributing to increased food insecurity—are catalysing efforts to identify more nutritious and affordable foods3. Consumption of dietary fibre can help to prevent cardiovascular disease, type 2 diabetes and obesity4–6. A substantial number of reports have explored the effects of dietary fibre on the gut microbial community7–9. However, the microbiome is complex, dynamic and exhibits considerable intra- and interpersonal variation in its composition and functions. The large number of potential interactions between the components of the microbiome makes it challenging to define the mechanisms by which food ingredients affect community properties. Here we address the question of how foods containing different fibre preparations can be designed to alter functions associated with specific components of the microbiome. Because a marked increase in snack consumption is associated with westernization, we formulated snack prototypes using plant fibres from different sustainable sources that targeted distinct features of the gut microbiomes of individuals with obesity when transplanted into gnotobiotic mice. We used these snacks to supplement controlled diets that were consumed by adult individuals with obesity or who were overweight. Fibre-specific changes in their microbiomes were linked to changes in their plasma proteomes indicative of an altered physiological state. Fibre snacks that target distinct features of the microbiomes of donors with obesity transplanted into gnotobiotic mice also lead to fibre-specific changes in the microbiome and physiology when used in controlled-diet human studies.

Precision medicine and sequence-based clinical diagnostics seek to predict disease risk or to identify causative variants from sequencing data. The Critical Assessment of Genome Interpretation (CAGI) is a community experiment consisting of genotype-phenotype prediction challenges; participants build models, undergo assessment, and share key findings. In the past, few CAGI challenges have addressed the impact of sequence variants on splicing. In CAGI5, two challenges (Vex-seq and MaPSY) involved prediction of the effect of variants, primarily single-nucleotide changes, on splicing. Although there are significant differences between these two challenges, both involved prediction of results from high-throughput exon inclusion assays. Here, we discuss the methods used to predict the impact of these variants on splicing, their performance, strengths, and weaknesses, and prospects for predicting the impact of sequence variation on splicing and disease phenotypes.

Neuroendocrine tumors (NET) and carcinomas (NEC) of the breast are rare diseases, but NEC has attracted attention in both cytopathology and surgical pathology because of its specific management and prognosis. Fine-needle aspiration biopsy (FNAB) cytology can make the diagnosis in many cases particularly with high-grade NEC, with definitive diagnosis based on histopathology and immunohistochemistry. This review describes the characteristics of the disease based on the WHO classification 2012 and recent literature and ­includes discussion related to the International Academy of Cytology Yokohama System of Reporting Breast FNAB ­cytology.

This article presents guidelines for imaging utilization in patients presenting with hearing loss or vertigo, symptoms that sometimes occur concurrently due to proximity of receptors and neural pathways responsible for hearing and balance. These guidelines take into account the superiority of CT in providing bony details and better soft-tissue resolution offered by MRI. It should be noted that a dedicated temporal bone CT rather than a head CT best achieves delineation of disease in many of these patients. Similarly, optimal assessment often requires a dedicated high-resolution protocol designed to assess temporal bone and internal auditory canals even though such a study will be requested and billed as a brain MRI. Angiographic techniques are helpful in some patients, especially in the setting of vertigo. The American College of Radiology Appropriateness Criteria are evidence-based guidelines for specific clinical conditions that are reviewed annually by a multidisciplinary expert panel. The guideline development and revision include an extensive analysis of current medical literature from peer reviewed journals and the application of well-established methodologies (RAND/UCLA Appropriateness Method and Grading of Recommendations Assessment, Development, and Evaluation or GRADE) to rate the appropriateness of imaging and treatment procedures for specific clinical scenarios. In those instances where evidence is lacking or equivocal, expert opinion may supplement the available evidence to recommend imaging or treatment.

Comorbid medical illnesses, such as obesity and diabetes, are associated with more severe COVID-19, hospitalization, and death. However, the role of the immune system in mediating these clinical outcomes has not been determined. We used multiparameter flow cytometry and systems serology to comprehensively profile the functions of T cells and antibodies targeting spike, nucleocapsid, and envelope proteins in a convalescent cohort of COVID-19 subjects who were either hospitalized (n = 20) or not hospitalized (n = 40). To avoid confounding, subjects were matched by age, sex, ethnicity, and date of symptom onset. Surprisingly, we found that the magnitude and functional breadth of virus-specific CD4+ T cell and antibody responses were consistently higher among hospitalized subjects, particularly those with medical comorbidities. However, an integrated analysis identified more coordination between polyfunctional CD4+ T cells and antibodies targeting the S1 domain of spike among subjects who were not hospitalized. These data reveal a functionally diverse and coordinated response between T cells and antibodies targeting SARS-CoV-2, which is reduced in the presence of comorbid illnesses that are known risk factors for severe COVID-19. CA extern

Comorbid medical illnesses, such as obesity and diabetes, are associated with more severe COVID-19, hospitalization, and death. However, the role of the immune system in mediating these clinical outcomes has not been determined. We used multi-parameter flow cytometry and systems serology to comprehensively profile the functions of T cells and antibodies targeting spike, nucleocapsid, and envelope proteins in a convalescent cohort of COVID-19 subjects who were either hospitalized (n=20) or not hospitalized (n=40). To avoid confounding, subjects were matched by age, sex, ethnicity, and date of symptom onset. Surprisingly, we found that the magnitude and functional breadth of virus-specific CD4 T cell and antibody responses were consistently higher among hospitalized subjects, particularly those with medical comorbidities. However, an integrated analysis identified more coordination between polyfunctional CD4 T-cells and antibodies targeting the S1 domain of spike among subjects that were not hospitalized. These data reveal a functionally diverse and coordinated response between T cells and antibodies targeting SARS-CoV-2 which is reduced in the presence of comorbid illnesses that are known risk factors for severe COVID-19. Our data suggest that isolated measurements of the magnitudes of spike-specific immune responses are likely insufficient to anticipate vaccine efficacy in high-risk populations.

Summary The recent advent of CRISPR and other molecular tools enabled the reconstruction of cell lineages based on induced DNA mutations and promises to solve the ones of more complex organisms. To date, no lineage reconstruction algorithms have been rigorously examined for their performance and robustness across dataset types and number of cells. To benchmark such methods, we decided to organize a DREAM challenge using in vitro experimental intMEMOIR recordings and in silico data for a C. elegans lineage tree of about 1,000 cells and a Mus musculus tree of 10,000 cells. Some of the 22 approaches submitted had excellent performance, but structural features of the trees prevented optimal reconstructions. Using smaller sub-trees as training sets proved to be a good approach for tuning algorithms to reconstruct larger trees. The simulation and reconstruction methods here generated delineate a potential way forward for solving larger cell lineage trees such as in mouse.

The pairing of CRISPR/Cas9-based gene editing with massively parallel single-cell readouts now enables large-scale lineage tracing. However, the rapid growth in complexity of data from these assays has outpaced our ability to accurately infer phylogenetic relationships. First, we introduce Cassiopeia—a suite of scalable maximum parsimony approaches for tree reconstruction. Second, we provide a simulation framework for evaluating algorithms and exploring lineage tracer design principles. Finally, we generate the most complex experimental lineage tracing dataset to date, 34,557 human cells continuously traced over 15 generations, and use it for benchmarking phylogenetic inference approaches. We show that Cassiopeia outperforms traditional methods by several metrics and under a wide variety of parameter regimes, and provide insight into the principles for the design of improved Cas9-enabled recorders. Together, these should broadly enable large-scale mammalian lineage tracing efforts. Cassiopeia and its benchmarking resources are publicly available at www.github.com/YosefLab/Cassiopeia.

Nifedipine (NF) is reported to have many beneficial effects in antihypertensive therapy. Recently, we found that NF induced lipid accumulation in renal tubular cells. Palmitic acid-induced renal lipotoxicity was found to be partially mediated by endoplasmic reticular (ER) stress, while it can also be elicited by NF in kidney cells, we examined the induction of suspected pathways in both in vitro and in vivo models. NRK52E cells cultured in high-glucose medium were treated with NF (30 &micro, M) for 24&ndash, 48 h. ER stress-induced lipotoxicity was explored by staining with thioflavin T and Nile red, transmission electron microscopy, terminal uridine nick-end labeling, and Western blotting. ER stress was also investigated in rats with induced chronic kidney disease (CKD) fed NF for four weeks. NF induced the production of unfolded protein aggregates, resulting in ER stress, as evidenced by the upregulation of glucose-regulated protein, 78 kDa (GRP78), activating transcription factor 6&alpha, (ATF6&alpha, ), C/EBP-homologous protein (CHOP), and caspases-12, -3, and -7. In vitro early apoptosis was more predominant than late apoptosis. Most importantly, ATF6&alpha, was confirmed to play a unique role in NF-induced ER stress in both models. CKD patients with hypertension should not undergo NF therapy. In cases where it is required, alleviation of ER stress should be considered to avoid further damaging the kidneys.

Summary Lowering total tau levels is an attractive therapeutic strategy for Alzheimer's disease and other tauopathies. High-throughput screening in neurons derived from human induced pluripotent stem cells (iPSCs) is a powerful tool to identify tau-targeted therapeutics. However, such screens have been hampered by heterogeneous neuronal production, high cost and low yield, and multi-step differentiation procedures. We engineered an isogenic iPSC line that harbors an inducible neurogenin 2 transgene, a transcription factor that rapidly converts iPSCs to neurons, integrated at the AAVS1 locus. Using a simplified two-step protocol, we differentiated these iPSCs into cortical glutamatergic neurons with minimal well-to-well variability. We developed a robust high-content screening assay to identify tau-lowering compounds in LOPAC and identified adrenergic receptors agonists as a class of compounds that reduce endogenous human tau. These techniques enable the use of human neurons for high-throughput screening of drugs to treat neurodegenerative disease., Highlights • Inducible, isogenic, and integrated Ngn2 iPSCs were differentiated to pure neurons • A simple and scalable two-step protocol ensures minimal differentiation variability • An HCS assay was established to screen for tau-lowering compounds in LOPAC • AR agonists were identified as a class of compounds that reduce human tau, Gan and colleagues developed a simple and scalable technology to generate a large quantity of homogeneous glutamatergic cortical neurons by engineering a neurogenin 2-expressing cassette to the AAVS1 locus of iPSCs. They developed a high-content screening assay and identified adrenergic receptor agonists as a class of compounds that lower endogenous human tau, a key pathogenic factor in Alzheimer's disease.

Perilla frutescens (L.) Britt Labiatae (Pl), known as “Zi-So” and widely grown in China (red cultivar, Rpl) and Japan (green cultivar, Gpl), is frequently served on “Sa-Shi-Mi” dishes. Considering ...

Memory impairment has been shown to be associated with glutamate (Glu) excitotoxicity, homocysteine (Hcy) accumulation, and oxidative stress. We hypothesize that Glu and Hcy could damage neuronal cells, while astaxanthin (ATX) could be beneficial to alleviate the adverse effects. Using PC12 cell model, we showed that Glu and Hcy provoked a huge amount of reactive oxygen species (ROS) production, causing mitochondrial damage at EC50 20 and 10 mm, respectively. The mechanisms of action include: (1) increasing calcium influx, (2) producing ROS, (3) initiating lipid peroxidation, (4) causing imbalance of the Bcl-2/Bax homeostasis, and (5) activating cascade of caspases involving caspases 12 and 3. Conclusively, the damages caused by Glu and Hcy to PC12 cells can be alleviated by the potent antioxidant ATX.

Taiwan produces large quantities of bananas in the southern area. Recently, due to the export quantity has been greatly reduced, in order to efficiently maintain the banana agriculture and economy, the development of alternate uses of bananas has become urgently in need. Bananas contain a fair amount of nutrients with low glycemic index. Currently, as the bread consumption is increasing, we tried to manufacture banana‐assorted breads. The desiccated powders of Musa sapientum var TC2‐425 Linn [(genomically, called as Musa (AAA) (MA)] and Musa basjoo “Nam Wa” (MB) were separately incorporated at 15%, 20%, and 25% (denoted as MA15‐MA25 and MB15‐MB25). Results indicated that MA exhibited higher contents of moisture, ash, crude protein, and lutein, while with lower crude fat, crude fibers, carbohydrate, sodium, total soluble sugars, and pectin. The contents of taste compounds (name, samples in decreasing order) were as follows: 5′‐CMP (MB25, MB20); 5′‐GMP (MA25, MA20); 5′‐AMP (MB25, MA15); 5′‐XMP (MA25, MA20); 5′‐IMP (MA25, MB20, MB25); and 5′‐UMP (MA20, MA25, MB20). Hedonic scoring (HS) indicated MA15, MA20, MB15, and MB20 were more acceptable. Textural profile analysis (TPA; for 0–6 days, only 0–4 days are shown) revealed that “flavor,” “mouthfeel,” “hardness,” “gumminess,” and “chewiness” were the determinant key roles. Conclusively, due to different chemical constituent of banana, different recipes must be considered. The bread acceptability is affected by the fermentative profile which in turn is governed by the contents of soluble sugars, pectin, taste compounds, and the overall activity of yeast cells., The bread acceptability is affected by the fermentative profile which in turn is governed by the contents of soluble sugars, pectin, and taste compounds. Hence, regarding the use of bananas in making the banana‐assorted breads, the recipes must be carefully designed by considering such factors.

To examine the contributions of impaired gut microbial community development to childhood undernutrition, we combined metabolomic and proteomic analyses of plasma samples with metagenomic analyses of fecal samples to characterize the biological state of Bangladeshi children with severe acute malnutrition (SAM) as they transitioned, after standard treatment, to moderate acute malnutrition (MAM) with persistent microbiota immaturity. Host and microbial effects of microbiota-directed complementary food (MDCF) prototypes targeting weaning-phase bacterial taxa underrepresented in SAM and MAM microbiota were characterized in gnotobiotic mice and gnotobiotic piglets colonized with age- and growth-discriminatory bacteria. A randomized, double-blind controlled feeding study identified a lead MDCF that changes the abundances of targeted bacteria and increases plasma biomarkers and mediators of growth, bone formation, neurodevelopment, and immune function in children with MAM. INTRODUCTION There is a dimension to post-natal human development that involves assembly of microbial communities in different body habitats, including the gut. Children with acute malnutrition have impaired development of their gut microbiota, leaving them with communities that appear younger (more immature) than those of chronologically age-matched healthy individuals. Current therapeutic foods given to children with acute malnutrition have not been formulated based on knowledge of how they affect the developmental biology of the gut microbiota. Moreover, they are largely ineffective in ameliorating the long-term sequelae of malnutrition that include persistent stunting, neurodevelopmental abnormalities, and immune dysfunction. RATIONALE Repairing microbiota immaturity and determining the degree to which such repair restores healthy growth requires identification of microbial targets that are not only biomarkers of community assembly but also mediators of various aspects of growth. Identifying ingredients in complementary foods, consumed during the transition from exclusive milk feeding to a fully weaned state, that increase the representation and expressed beneficial functions of growth-promoting bacterial taxa in the developing microbiota could provide an effective, affordable, culturally acceptable, and sustainable approach to treatment. RESULTS Metabolomic and proteomic analyses of serially collected plasma samples were combined with metagenomic analyses of serially collected fecal samples from Bangladeshi children with severe acute malnutrition (SAM) treated with standard therapy. The results provided a readout of their biological features as they transitioned from SAM to a state of persistent moderate acute malnutrition (MAM) with accompanying persistent microbiota immaturity. Significant correlations were identified between levels of plasma proteins, anthropometry, plasma metabolites, and the representation of bacteria in their microbiota. Gnotobiotic mice were subsequently colonized with a defined consortium of bacterial strains that represent various phases of microbiota development in healthy Bangladeshi children. Administration of different combinations of Bangladeshi complementary food ingredients to colonized mice and germ-free controls revealed diet-dependent increases in the abundance and changes in the metabolic activities of targeted weaning-phase strains as well as diet- and colonization-dependent augmentation of growth-promoting host signaling pathways. Host and microbial effects of microbiota-directed complementary food (MDCF) prototypes were subsequently examined in gnotobiotic mice colonized with immature microbiota from children with post-SAM MAM and in gnotobiotic piglets colonized with a defined consortium of targeted age- and growth-discriminatory taxa. A randomized, double-blind study of standard therapy versus various MDCF prototypes emerging from these preclinical models, conducted in Bangladeshi children with MAM, identified a lead MDCF that increased levels of biomarkers and mediators of growth, bone formation, neurodevelopment, and immune function toward a state resembling healthy children. Using an approach inspired by statistical methods applied to financial markets, we show in the accompanying paper by Raman et al. that this lead MDCF was most effective in repairing the microbiota. CONCLUSION These findings demonstrate the translatability of results obtained from pre-clinical gnotobiotic animal models to humans, directly support the hypothesis that healthy microbiota development is causally linked to healthy growth, illustrate an approach for treating childhood undernutrition, and with the capacity to deliberately reconfigure immature microbiota, suggest a means to decipher how elements of the gut microbial community operate to regulate various host systems involved in healthy growth. Overview of therapeutic food discovery and testing. The approach used for integrating preclinical gnotobiotic animal models with human studies to understand the contributions of perturbed gut microbiota development to childhood malnutrition and to identify MDCFs.

Accurate interpretation of genomic variants that alter RNA splicing is critical to precision medicine. We present a computational framework, PEPSI (Prediction of variant Effect on Percent Spliced In), that predicts the splicing impact of coding and noncoding variants for the Fifth Critical Assessment of Genome Interpretation (CAGI5) “Vex-seq” challenge. PEPSI is a random forest regression model trained on multiple layers of features associated with sequence conservation and regulatory sequence elements. Compared to other splicing defect prediction tools from the literature, our framework integrates secondary structure information in predicting variants that disrupt splicing regulatory elements (SREs). We applied our model to classify splice-disrupting variants among 2,094 single nucleotide polymorphisms (SNPs) from the Exome Aggregation Consortium (ExAC) using model-predicted changes in percent spliced in (ΔPSI) associated with tested variants. Benchmarking our model against widely used state-of-the-art tools, we demonstrate that PEPSI achieves comparable performance in terms of sensitivity and precision. Moreover, we also show that using secondary structure context can help resolve several cases where changes in the counts of SREs do not correspond with the directionality of ΔPSI measured for tested variants. Our tool is freely available to researchers at: https://github.com/rwang916/PEPSI

Seven kinds of indigenous fruits and five imported fruits were compared for their "health quality." Methods including the calorific, antioxidant, and ion ratios were carried out. Results indicated the order of content (in mg/100g) was: Ca(2+) , Murcott orange (218.2) > Kiwifruit (200.0) > pineapple (138.5) > Golden kiwi (117.6); Mg(2+) , Pitaya (192.2), banana (88.0), Kiwifruit (63.4), and Golden kiwi (58.4); Zn(2+) , Pitaya (19.53) > pear (10.8) > Kiwifruit (6.09) > Irwin mango (4.58). Cu(2+) , Kiwifruit (0.70) > Red globe grape (0.67) > Golden kiwi (0.65) > Irwin mango (0.42) ≈ Pitaya (0.40). In terms of ion ratio, Pitaya showed Zn(2+) /Cu(2+) (48.8), Mg(2+) /Ca(2+) (6.7) and uniquely possessed selenium 0.002 mg/100 g; for pear, Zn(2+) /Cu(2+) = 37.2, while Kyoho grape, Red globe grape, and Golden kiwi revealed extremely high Fe(2+) /(Co(2+) +Ni(2+) ) ratios. On the other hands, Irwin mango and Pitaya astonishingly contained huge amount of inositol, reaching 3523.2 mg/100 g and 1998.7 mg/100 g, respectively. To evaluate the "health quality" of fruits, an overall ranking method by combining (a) the Function-directed Caloric Antioxidant Density (CAD) and (b) the ion ratio was developed. The finalized ranking of these selected fruits was: Pitaya > cherry > Irwin mango > Murcott orange = pineapple > banana > Golden kiwi > pear > Kiwifruit > Red globe grape > apple > Kyoho grape. Conclusively, this evaluation method is novel, contemporary and scientific, which could more clearly assess the "health quality" of fruits in view of nutritional, calorific, and antioxidant balance.

Rhizoma Alpinia officinarum (Hance) Farw, Zingiberaceae (AO), a ginger family herb exhibiting stimulant and a carminative bioactivity, is widely used in European and Asian countries as spicy condiment and medicinal uses. Allyl isothiocyanate (AITC) is the main pungent taste of native Wasabi (Wasabia japonica). The cytotoxicity of AITC has been implicated in thymus, adrenals, and white blood cells. Considering food safety, apparently a safer substitute for wasabi is worthy commercialized. Previously, we found AO crude paste to be rather feasible for use as a "Wasabi-substitute" in fresh meat and cold salads. A process linking cold ethyl acetate (EtAc) extraction with silica gel adsorption and reversed phase high-performance liquid chromatography (RP-HPLC) (mobile phase, 75% methanol) was used to isolate galangal acetate, the Wasabi-like taste constituent. AO contained abundant galangal acetate (3.84 ± 0.07%) compared to A. galangal (0.57 ± 0.16%), and as already confirmed by thin layer chromatography (TLC), gas chromatography (GC)/mass spectrometry (MS)/MS and Nuclear Magnetic Resonance Spectroscopy (NMR), galangal acetate was particularly thermally labile. The steam distilled essential oil (SDEO) of AO (0.14% on wet basis) contained 80 compounds (number of component, %): monoterpene hydrocarbon (21, 13.83%); oxygenated monoterpene (17, 27.08%); sesquiterpene hydrocarbon (20, 31.03%), and oxygenated sesquiterpene (20, 21.85%), respectively. However, no spicy wasabi-like constituent remained in SDEO. Alternatively, n-hexane, EtAc, and methanol extracts of AO all showed potent DPPH- and superoxide anion-scavenging activity. Conclusively, SDEO although contains 80 volatiles, galangal acetate is absent due to thermal instability. Galangal acetate exhibits pleasant "Wasabi-like taste" for which we have successively developed an integrated process for mass production.

Treatment of ovarian cancer, a leading cause of gynecological malignancy, has good initial efficacy with surgery and platinum/taxane-based chemotherapy, but poor long-term survival in patients. Inferior long-term prognosis is attributed to intraperitoneal spreading, relapse and ineffective alternate therapies. Adoptive cell therapy is promising for tumor remission, although logistical concerns impede widespread implementation. In this study, healthy PBMCs were used to examine the immune response in a mouse model with human ovarian cancer, where natural killer (NK) cells were found to be the effector cells that elicited an anti-tumor response. Presence of tumor was found to stimulate NK cell expansion in mice treated intraperitoneally with PBMC+Interleukin-2 (IL-2), as compared to no expansion in non-tumor-bearing mice given the same treatment. PBMC+IL-2 treated mice exhibiting NK cell expansion had complete tumor remission. To validate NK cell mediated anti-tumor response, the intratumoral presence of NK cells and their cytotoxicity was confirmed by immunohistochemistry and granzyme activity of NK cells recovered from the tumor. Collectively, this study highlights the significance of NK cell-cytotoxic response to tumor, which may be attributed to interacting immune cell types in the PBMC population, as opposed to clinically used isolated NK cells showing lack of anti-tumor efficacy in ovarian cancer patients.

Hericium erinaceus (Bulliard: Fries) Persoon (HE), synonymously called Monkey Head mushroom or Yamabushitake, contains a huge amount of nutrients, in particular, the erinacines. This study was carried out with the aim to uncover whether erinacines can play a role as a neuroprotective adjuvant and what the related mechanism of action could be. A novel medium was developed to significantly enhance the productivity of biomass as well as erinacines to reach 16.8 g/L and 225 ± 54 mg/L, respectively. The ethanolic extract of HE (HEE) was tested on PC-12 cell line for its protective effect against the 20 mM glutamate-insult (GluI). The parameters affected were (control vs. GluI): glutathione (8.9 ± 0.8 vs. 2.5 ± 0.7 nmol/mg protein); glutathione peroxidase (11.6 ± 1.6 vs. 28.2 ± 3.2 mU/mg protein); glutathione reductase (9.2 ± 0.8 vs. 2.3 ± 0.4 mU/mg-protein); Ca2+-influx (115 ± 16 vs. 360 ± 23 nmol/L); reactive oxygen species (100 ± 3 vs. 140 ± 7%); superoxide dismutase (56.5 ± 6.3 vs. 23.2 ± 4.2 U/mg-protein); H2O2 (8.2 ± 0.7 vs. 20.4 ± 3.5 nmol/mg-protein); and TBARs (MDA) (8.1 ± 0.3 vs. 13.3 ± 2.5 μmol/μg-protein) (p

Quantitative assessments of the capacity of marine reserves to restore historical fish body-size distributions require extensive repeated sampling to map the phenotypic responses of target populations to protection. However, the "no take" status of marine reserves oftentimes precludes repeated sampling within their borders and, as a result, our current understanding of the capacity of marine reserves to restore historical body-size distributions remains almost entirely reliant on independent, static visual surveys. To overcome this challenge, we promote the application of a traditional fisheries tool known as a "back-calculation", which allows for the estimation of fish body lengths from otolith annuli distances. This practical application was pursued in this study, using data collected in five marine reserves and adjacent fished reefs in the Philippines, to investigate spatiotemporal disparities in length-at-age of the brown surgeonfish, Acanthurus nigrofuscus. The spatial component of our analyses revealed that 1) A. nigrofuscus were phenotypically similar between marine reserves and fished reefs during their early life history; 2) marine reserve and fished reef populations diverged into significantly different length-at-age morphs between ages three and six, in which protected fish were predominantly larger than conspecifics in fished reefs; and 3) A. nigrofuscus returned to a state of general phenotypic similarity during later life. The temporal component of our analyses revealed that younger generations of A. nigrofuscus exhibited significant, positive year effects that were maintained until age eight, indicating that, within the significant age cohorts, younger generations were significantly larger than older generations.

Introduction Therapy of marrow borne disease such as leukemias or myelomas require the eradication of tumor cells that are resident in the bone marrow. Cell based therapies are increasingly a promising modality toward potential complete cures of such marrow malignancies. In particular, natural killer (NK) cell therapeutics are an attractive therapeutic modality due to their inherent ability to broadly detect and kill tumors. However, for NK cells to kill the marrow resident blasts, they must traffic to the bone marrow. Key for marrow homing is the proper fucosylation of extracellular proteins (such as e.g. P-selectin glycoprotein ligand-1, PSGL) on NK cells which is necessary for E-selectin binding. PM21 particles are plasma membrane particles prepared from CSTX002 cell line (K562 cells modified to express membrane bound IL-21 [mbIL21] and 4-1bbl, K562.mbIL21.41BBL) and can be used to potently stimulate expansion of NK cells that have high anti-tumor potency, similarly as to the expansion with CSTX002 feeder cells that have mbIL21 stimulation. Shown herein is that NK cells stimulated with PM21 particles have increased fucosylation of E-selectin binding ligands and have increased homing to the bone marrow in mice. Methods NK cells were expanded from PBMCs obtained from healthy donors by stimulating with PM21 particles (200 µg/mL) in SCGM media supplemented with 10% FBS and IL-2 (100 U/mL). PM21 particles were prepared from CSTX002 cell line as previously described. The fucosylation status of E-selectin ligands on expanded NK cells were analyzed by flow cytometry with the HECA452 mAb that only binds the fucosylated forms. ChIP-seq analysis was performed on NK cells expanded from feeder cell co-cultures with CSTX002 and by probing with anti-STAT3. For physiological trafficking studies, whole PBMCs were pre-activated in vitro for two days with PM21 particles (200 µg/mL) and then injected ip into NSG mice and treated or not with PM21-particles (2x/weekly). Organs were harvested 16 days later and analyzed by flow cytometry for the presence of human NK cells. Results NK cells expanded using PM21 or other stimulation methods were comparatively analyzed for fucosylation status by staining with HECA452. The NK cells stimulated with mbIL21, either through PM21 or CSTX002 feeder cells, were found to have highest HECA452 staining compared NK cells stimulated with IL-2, soluble IL-21, soluble 4-bbl, unmodified K562, or K562 modified with only mbIL21 (no 4-1bbl). To determine transcriptional changes responsible for the apparent increase in fucosylation, anti-STAT3 ChIP-seq analysis was performed on naïve NK cells or those expanded with CSTX002. STAT3 is a transcription factor that is activated upon IL-21 stimulation. The anti-STAT3 ChIP-seq analysis elucidated that CSTX002 stimulated NK cells have enhanced STAT3 binding to the FUT7 gene and that FUT7 expression increased. To determine if stimulation with PM21 particles can enhance homing of NK cells to bone marrow, NSG mice were injected ip with NK cells stimulated in vitro for 2 days with PM21, and then the mice were treated with or without PM21 particles (ip, 800 µg, 3x per week). Analysis of the bone marrow from sacrificed animals showed that a greater amount of human NK cell were present in the marrow in animals treated with ip doses of PM21 particles. Conclusions NK cells expanded with PM21 particles had increased fucosylation of likely E-selecting ligands. This higher degree of fucosylation is likely due to the enhanced expression of fucosyl transferase 7, shown bye ChIP-seq analysis of CSTX002 stimulated NK cells. In vivo studies show that stimulation with PM21 does allow for increased trafficking of NK cells to the bone marrow. Disclosures Oyer: Kiadis Pharma: Equity Ownership, Patents & Royalties. Igarashi:Kiadis Pharma: Employment, Equity Ownership, Patents & Royalties. Altamore:Kiadis Pharma: Equity Ownership, Patents & Royalties. Lee:Kiadis Pharma: Consultancy, Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Patents & Royalties, Research Funding. Copik:Kiadis Pharma: Equity Ownership, Patents & Royalties, Research Funding.

Differences in the expression of genes and their splice isoforms across human tissues are fundamental factors to consider for therapeutic target evaluation. To this end, we conducted a transcriptome-wide survey of tissue-specific gene expression and splicing events in the unprecedented collection of 8527 high-quality RNA-seq samples from the GTEx project, covering 36 human peripheral tissues and 13 brain subregions. We derived a weighted tissue-specificity scoring scheme accounting for the similarity of related tissues and inherent variability across individual samples. We showed that ~50.6% of all annotated human genes show tissue-specific expression, including many low abundance transcripts vastly underestimated by previous array-based expression atlases. As utilities for drug discovery, we demonstrated that tissue-specificity is a highly desirable attribute of validated drug targets and tissue-specificity can be used to prioritize disease-associated genes from genome-wide association studies (GWAS). Using brain striatum-specific gene expression as an example, we provided a template to leverage tissue-specific gene expression to identify novel therapeutic targets. Mining of tissue-specific splicing further reveals new opportunities for tissue-specific targeting. Thus, the high quality transcriptome atlas provided by the GTEx is an invaluable resource for drug discovery and systematic analysis anchored on the human tissue specific gene expression provides a promising avenue to identify novel therapeutic target hypotheses.

To show and rationalize the confounding effects on the rotational/oscillatory rheology of surface active impurities in commercial protein formulations such as bovine serum albumin, BSA. Bulk and interfacial rotational/oscillatory rheology were used to study the viscosity, complex viscosity, storage/elastic modulus, G’ and loss/viscous modulus, G”, as a function of time of aqueous formulations of BSA and their purified components. Viscosity/time profiles at steady shear for different commercial BSA products and lots showed viscosity increase, decrease and time-independent profiles at low shear rates. All lots showed shear thinning. BSA monomer and dimers/aggregates, in general, showed similar profiles. Addition of low levels of surfactant or high shear rates rendered all solutions to be Newtonian-like. Interfacial viscosity studies paralleled those on the rotational rheometer. G’ > G” with viscosity increase and G’

Human fishing effort is size-selective, preferentially removing the largest individuals from harvested stocks. Intensive, size-specific fishing mortality induces directional shifts in phenotypic frequencies towards the predominance of smaller and earlier-maturing individuals, which are among the primary causes of declining fish biomass. Fish that reproduce at smaller size and younger age produce fewer, smaller, and less viable larvae, severely reducing the reproductive capacity of harvested populations. Marine protected areas (MPAs) are extensively utilized in coral reefs for fisheries management, and are thought to mitigate the impacts of size-selective fishing mortality and supplement fished stocks through larval export. However, empirical evidence of disparities in fitness-relevant phenotypes between MPAs and adjacent fished reefs is necessary to validate this assertion. Here, we compare key life-history traits in three coral-reef fishes (Acanthurus nigrofuscus, Ctenochaetus striatus, and Parupeneus multifasciatus) between MPAs and fished reefs in the Philippines. Results of our analyses support previous hypotheses regarding the impacts of MPAs on phenotypic traits. Asymptotic length (Linf) and growth rates (K) differed between conspecifics in MPAs and fished reefs, with protected populations exhibiting phenotypes that are known to confer higher fecundity. Additionally, populations demonstrated increases in length at 50% maturity (L50) inside MPAs compared to adjacent areas, although age at 50% maturity (A50) did not appear to be impacted by MPA establishment. Shifts toward advantageous phenotypes were most common in the oldest and largest MPAs, but occurred in all of the MPAs examined. These results suggest that MPAs may provide protection against the impacts of size-selective harvest on life-history traits in coral-reef fishes.

Summary Human astrocytes network with neurons in dynamic ways that are still poorly defined. Our ability to model this relationship is hampered by the lack of relevant and convenient tools to recapitulate this complex interaction. To address this barrier, we have devised efficient coculture systems utilizing 3D organoid-like spheres, termed asteroids, containing pre-differentiated human pluripotent stem cell (hPSC)-derived astrocytes (hAstros) combined with neurons generated from hPSC-derived neural stem cells (hNeurons) or directly induced via Neurogenin 2 overexpression (iNeurons). Our systematic methods rapidly produce structurally complex hAstros and synapses in high-density coculture with iNeurons in precise numbers, allowing for improved studies of neural circuit function, disease modeling, and drug screening. We conclude that these bioengineered neural circuit model systems are reliable and scalable tools to accurately study aspects of human astrocyte-neuron functional properties while being easily accessible for cell-type-specific manipulations and observations., Highlights • Novel three-dimensional “asteroid” coculture system is described • Complex human astrocyte-specific morphology is generated in vitro • This system produces rapid and tight association of astrocytes with synapses, In this article, Krencik and colleagues show that high-density cocultures of pre-differentiated human astrocytes with induced neurons, from pluripotent stem cells, elicit mature characteristics by 3–5 weeks. This provides a faster and more defined alternative method to organoid cultures for investigating human neural circuit function.

Natural killer (NK) cell immunotherapy as a cancer treatment shows promise, but expanding NK cells consistently from a small fraction (∼5%) of peripheral blood mononuclear cells (PBMCs) to therapeutic amounts remains challenging. Most current ex vivo expansion methods use co-culture with feeder cells (FC), but their use poses challenges for wide clinical application. We developed a particle-based NK cell expansion technology that uses plasma membrane particles (PM-particles) derived from K562-mbIL15-41BBL FCs. These PM-particles induce selective expansion of NK cells from unsorted PBMCs, with NK cells increasing 250-fold (median, 35; 10 donors; range, 94 to 1492) after 14 days of culture and up to 1265-fold (n = 14; range, 280 to 4426) typically after 17 days. The rate and efficiency of NK cell expansions with PM-particles and live FCs are comparable and far better than stimulation with soluble 41BBL, IL-15, and IL-2. Furthermore, NK cells expand selectively with PM-particles to 86% (median, 35; range, 71% to 99%) of total cells after 14 days. The extent of NK cell expansion and cell content was PM-particle concentration dependent. These NK cells were highly cytotoxic against several leukemic cell lines and also against patient acute myelogenous leukemia blasts. Phenotype analysis of these PM-particle–expanded NK cells was consistent with an activated cytotoxic phenotype. This novel NK cell expansion methodology has promising clinical therapeutic implications.

Both recombinant and natural human IgG2 antibodies have several different disulfide bond isoforms, which possess different global structures, thermal stabilities, and biological activities. A detailed mapping of the structural difference among IgG2 disulfide isoforms, however, has not been established. In this work, we employed hydrogen/deuterium exchange mass spectrometry to study the conformation of three major IgG2 disulfide isoforms known as IgG2-B, IgG2-A1, and IgG2-A2 in two recombinant human IgG2 monoclonal antibodies. By comparing the protection factors between amino acid residues in isoforms B and A1 (the classical form), we successfully identified several local regions in which the IgG2-B isoform showed more solvent protection than the IgG2-A1 isoform. On the basis of three-dimensional structural models of IgG2, these identified regions were located on the Fab domains, close to the hinge, centered on the side where the two Fab arms faced each other in spatial proximity. We speculated that in the more solvent-protected B isoform, the two Fab arms were brought into contact by the nonclassical disulfide bonds, resulting in a more compact global structure. Loss of Fab domain flexibility in IgG2-B could limit its ability to access cell-surface epitopes, leading to reduced antigen binding potency. The A2 isoform was previously found to have disulfide linkages similar to those of the classical A1 isoform, but with different biophysical behaviors. Our data indicated that, compared to IgG2-A1, IgG2-A2 had less solvent protection in some heavy-chain Fab regions close the hinge, suggesting that the A2 isoform had more flexible Fab domains.

Nitric oxide (NO), an essential agent of the innate immune system, exhibits multi-mechanistic antimicrobial activity. Previously, NO-releasing nanoparticles (NO-np) demonstrated increased antimicrobial activity when combined with glutathione (GSH) due to formation of S-nitrosoglutathione (GSNO), a transnitrosylating agent. To capitalize on this finding, we incorporated the thiol-containing ACE-inhibitor, captopril, with NO-np to form SNO-CAP-np, nanoparticles that both release NO and form S-nitrosocaptopril. In the presence of GSH, SNO-CAP-np demonstrated increased transnitrosylation activity compared to NO-np, as exhibited by increased GSNO formation. Escherichia coli and methicillin-resistant Staphylococcus aureus were highly susceptible to SNO-CAP-np in a dose-dependent fashion, with E. coli being most susceptible, and SNO-CAP-np were nontoxic in zebrafish embryos at translatable concentrations. Given SNO-CAP-np's increased transnitrosylation activity and increased E. coli susceptibility compared to NO-np, transnitrosylation rather than free NO is likely responsible for overcoming E. coli 's resistance mechanisms and ultimately killing the pathogen. From the Clinical Editor This team of authors incorporated the thiol-containing ACE-inhibitor, captopril, into a nitric oxide releasing nanoparticle system, generating nanoparticles that both release NO and form S-nitrosocaptopril, with pronounced toxic effects on MRSA and E. coli in the presented model system.

Hepatic CYP2B metabolizes cyclophosphamide (CP) into acrolein and phosphoramide mustard, which are the ultimate toxic teratogenic compounds. To determine whether folic acid (FA), resveratrol (RV) and vitamin E (vitE) could prevent these adverse effects, we carried out this study with CP at 20–60 nM. CP at 40 nM yielded maximum malformation (58.3 ± 4.2%) with a minimum mortality rate (58.3 ± 4.5%) and severe neural tube defects (NTDs). CP (40 nM) was co-administered with nutraceutics (each at 0.2 and 2.0 μM) applied to day-zero chicken embryo models (CEMs) and the biochemical parameters in brains were examined. CP upregulated (vs. control) hydrogen peroxide (HPO) (1.75/1.05 μmol per μg protein), hif-1α gene (1.5 fold), and HIF-1α protein (1.87 fold). CP downregulated glutathione (3.0/5.2 μmol per μg protein), cytochrome c oxidase (CCO) (10/22 U per mg protein) and ATP production (0.52 ± 0.7/1.23 ± 0.4 pmol per μg protein) (p 20 U per mg protein, and ATP to >1.2 pmol per mg protein. As a consequence, the intracranial hemorrhage was completely alleviated. Interestingly, vitE at 2.0 μM slightly upregulated the hif-1α gene, and suppressed GSH, CCO, and ATP production, which was suggested to be due to limited permeability through the blood brain barrier and the formation of tocopheryl free radicals. Conclusively, CP tends to elicit intracranial hemorrhaging via the mitochondrial ROS-hif-1α-ATP depletion pathway. For the prevention of such teratogenicity, FA and RV at 2.0 μM are more effective than vitE.

Gallic acid (3,4,5-trihydroxybenzoic acid) (GA) occurs in many plants. The adverse effects of GA are seldom cited. GA (6–14 μM) provoked the hemorrhagic liposis of the cervical muscles and intracranial hemorrhage. The cause of these pathological events and the method for prevention are still lacking. Using the chicken embryo model and some selected nutraceutics such as folate, glutathione (GSH), N-acetylcysteine, and vitamin E (Vit E), we carried out this study. Results revealed that the action mechanism of GA involved (i) inducing hypoxia with upregulated gene hif-1α and downregulated ratio vegf-r2/vegf-a, leading to dys-vascularization and myopathy; (ii) impairing cytochrome c oxidase; (iii) stimulating creatine kinase and lactate dehydrogenase release; (iv) eliciting carnitine accumulation and liposis via downregulating gene CPT1; (v) suppressing superoxide dismutase and stimulating NO, H2O2, and malondialdehyde; and (vi) depleting erythrocytic and tissue GSH, resulting in hemorrhage. When both Vit E and GSH were applied to the day 1 chicks, a better alleviation effect was revealed. Conclusively, GA potentially exhibits adverse effect by eliciting hemorrhagic liposis of cervical muscles and cerebral hemorrhage. Supplementation with GSH, Vit E, and N-acetylcysteine is able to ameliorate these adverse effects, warranting the importance of restricting the clinical phytotherapeutic doses of GA and related compounds. Copyright © 2014 John Wiley & Sons, Ltd.

In the Southern Ocean, it is still not certain that overall krill biomass may decline because of drastic increase in pCO2, and consequent decline in pH. However, there is evidence that ecological vacuums created by krill population collapses caused by ecosystem shifts in Western Antarctic Peninsula (WAP) region led to replacement of Antarctic krill Euphausia superba by soft-bodied salps Salpa thomsoni. There is yet another questionable hypothesis that by the end of 21st century, ocean acidification stress, coupled with thermal increase, may synergistically induce physiologically critical stress to Antarctic krill in some areas of the Southern Ocean, egg development of krill may drastically decrease and in the 23rd century krill may even become extinct. I have earlier reported on normal krill egg development in relation to thermal change and high pressure (George and Stromberg, 1985). Recent experiments on krill development under different pCO2 conditions by Kawaguchi et al. (2011, 2013) suggest that we may witness 20 to 70 % reduction in Antarctic Krill by 2100 as direct consequence of pH decline. Such a scenario may lead to demise of krill-eating top-predators like baleen whales, seals and different species of Antarctic penguin populations. We now know that Adelaide penguins are decreasing in Bransfield Strait region off of the Western Antarctic Peninsula but increasing in Ross Sea region. Such a shift in breeding colonies moving from northern to southern WAP region and Ross Sea areas is not attributed to any decline in krill biomass but recent decadal melting of sea-ice as documented by remote sensing (George and Hayden, 2017). In this paper the main focus revolves around implications of changing chemistry of the Southern Ocean caused by absorption of anthropogenic carbon dioxide.

The Sargasso Sea has been a ocean life habitat for millions of years, yet accurate assessment of the boundary area and detection of these relatively small sea surface features using Landsat series and Moderate resolution Imaging Spectroradiometer (MODIS) instruments have been found to have difficulty or even impossible due to lack of spatial resolution, coverage, recurring observance, and algorithm limitations to identify pelagic species of Sargassum. Sargassum rafts tend to be elongated, curved in the upwind direction and warmer than the surrounding ocean surface. Long weed ‘trails’ extending upwind from the rafts are evidence of plants dropping out and being left behind. Satellite data utilizing a simple ocean color indexes such as the floating algae Index and Normalized Differential Vegetation Index (NDVI) have been established to detect floating algae in open environments using MODIS instruments. Floating Algae Index (FAI) has shown advantages over the traditional NDVI and Enhanced Vegetation Index (EVI) because FAI is less sensitive to changes in environmental and observing conditions (aerosol type and thickness, solar/viewing geometry, and sun glint) and can see through thin clouds. The baseline subtraction method provides a simple yet effective means for atmospheric correction. The algorithms assisted in identifying the boundary area of the Sargasso Sea and the path of this floating alga past Cape Hatteras into the Atlantic Ocean. Due to the fact that similar spectral bands are available on many existing and planned satellite sensors such as Landsat series observations satellites, the NDVI and FIA concept was extendable to establish a long-term record of these ecologically biological dependent ocean plants.

Agonism of cell surface receptors by monoclonal antibodies is dependent not only on its ability to bind the target, but also to deliver a biological signal through receptors to the cell. Immunoglobulin G2 antibodies (IgG2s) are made up of a mixture of distinct isoforms (IgG2-A, -B and A/B), which differ by the disulfide connectivity at the hinge region. When evaluating panels of agonistic antibodies against CD200 receptor (CD200R) or βklotho receptor (βklotho), we noticed striking activity differences of IgG1 or IgG2 antibodies with the same variable domains. For the CD200R antibody, the IgG2 antibody demonstrated higher activity than the IgG1 or IgG4 antibody. More significantly, for βklotho, agonist antibodies with higher biological activity as either IgG2 or IgG1 were identified. In both cases, ion exchange chromatography was able to isolate the bioactivity to the IgG2-B isoform from the IgG2 parental mixture. The subclass-related increase in agonist activity was not correlated with antibody aggregation or binding affinity, but was driven by enhanced avidity for the CD200R antibody. These results add to the growing body of evidence that show that conformational differences in the antibody hinge region can have a dramatic impact on the antibody activity and must be considered when screening and engineering therapeutic antibody candidates. The results also demonstrate that the IgG1 (IgG2-A like) or the IgG2-B form may provide the most active form of agonist antibodies for different antibodies and targets.

Human IgG2 consists of disulfide-mediated structural isoforms, classified by the number of Fab arms disulfide-linked to the heavy chain hinge. In the IgG2-B isoform, both Fab arms are linked to the hinge region, and in IgG2-A, neither Fab arm are linked to the hinge. IgG2-A/B is a hybrid between these two forms, with only one Fab arm disulfide-linked to the hinge. Within each of these isoform types are subtypes, with subtle disulfide-linkage differences. Here we explored the structural basis for the A1 and A2 isoform subtypes. Whereas A1 isoform converts into the A/B and B isoforms under mild redox conditions, A2 does not. Characterization of the disulfide connectivities of A2 isoform revealed a similar structure to A1 isoform, with parallel inter heavy chain disulfide linkages in the hinge region. However, the hinge disulfides in A2 isoform were resistant to reduction under conditions where A1 isoform hinge disulfides became reduced and they required thermal treatment (>55°C) to obtain thiol-dependent disulfide reduction. Structural analysis of the hinge region indicated that the protected disulfides were restricted to cysteines 219 and 220 of the upper hinge. Disruption of the upper hinge through insertion mutagenesis eliminated A2 isoform behavior. 1H NMR studies showed that the A1 isoform Fc glycan was more dynamic than that on A2 isoform and showed some other conformational differences. Results point to an IgG2-A2 upper hinge region that is more akin to the interior of a globular protein than the flexible hinge region expected on an IgG.

Whereas DC have increasingly been recognized for their role in activating the inflammatory cascades during IRIs, the mechanisms by which oxidative stress enhances DC activation remain to be explored. We examined the role of oxidative stress on two important features of DC: T cell activation and trafficking. Bone marrow-derived OS-DC were compared with untreated DC. DC exposed to oxidative stress augmented allogeneic T cell proliferation and showed increased migration in a chemotaxis chamber. These results were confirmed by using hypoxanthine and xanthine oxidase as another inducer of oxidative stress. We used OT-II and OT-I mice to assess the effect of oxidative stress on DC activation of OVA-specific CD4+ and CD8+ T cells, respectively. Oxidative stress increased DC capacity to promote OVA-specific CD4+ T cell activity, demonstrated by an increase in their proliferation and production of IFN-γ, IL-6, and IL-2 proinflammatory cytokines. Whereas oxidative stress increased the DC ability to stimulate IFN-γ production by OVA-specific CD8+ T cells, cellular proliferation and cytotoxicity were not affected. Compared with untreated DC, oxidative stress significantly reduced the capacity of DC to generate Tregs, which were restored by using anti-IL-6. With regard to DC trafficking, whereas oxidative stress increased DC expression of p-Akt and p-NF-κB, targeting PI3Kγ and NF-κB pathways abrogated the observed increase in DC migration. Our data propose novel insights on the activation of DC by oxidative stress and provide rationales for targeted therapies, which can potentially attenuate IRI.

As a consequence of intense fishing pressure, fished populations experience reduced population sizes and shifts in body size toward the predominance of smaller and early maturing individuals. Small, early-maturing fish exhibit significantly reduced reproductive output and, ultimately, reduced fitness. As part of resource management and biodiversity conservation programs worldwide, no-take marine protected areas (MPAs) are expected to ameliorate the adverse effects of fishing pressure. In an attempt to advance our understanding of how coral reef MPAs meet their long-term goals, this study used visual census data from 23 MPAs and fished reefs in the Philippines to address three questions: (1) Do MPAs promote shifts in fish body size frequency distribution towards larger body sizes when compared to fished reefs? (2) Do MPA size and (3) age contribute to the efficacy of MPAs in promoting such shifts? This study revealed that across all MPAs surveyed, the distribution of fishes between MPAs and fished reefs were similar, however, large-bodied fish were more abundant within MPAs, along with small, young-of-the-year individuals. Additionally, there was a significant shift in body size frequency distribution towards larger body sizes in 12 of 23 individual reef sites surveyed. Of 22 fish families, eleven demonstrated significantly different body size frequency distributions between MPAs and fished reefs, indicating that shifts in the size spectrum of fishes in response to protection are family-specific. Family-level shifts demonstrated a significant, positive correlation with MPA age, indicating that MPAs become more effective at increasing the density of large-bodied fish within their boundaries over time.

Recent animal and human studies have demonstrated the importance of the ROCK (RhoA/Rho-associated kinase) pathway in IsST (ischaemic stroke). Whether the genetic variation within ROCK-associated genes modulates the risk of IsST remains elusive. The association between 66 tSNPs [tagging SNPs (single nucleotide polymorphisms)] of three ROCK-associated genes [ROCK1, ROCK2 and ARHGEF10 (Rho guanine-nucleotide-exchange factor 10)] and the incidence of IsST was investigated in 23294 Caucasian female participants of the prospective WGHS (Women's Genome Health Study). All were free of known cancer and cardiovascular disease at baseline. During a 15-year follow-up period, 323 participants developed their first ever IsST. Multivariable Cox regression analysis was performed to investigate the relationship between genotypes and risk of IsST assuming an additive genetic model. Haplotype-block analysis was also performed. A total of ten tSNPs were associated with the risk of IsST (three in ARHGEF10 and seven in ROCK1; P

Individual and sexual partner characteristics may increase the risk of abnormal cervical cytology among women in human immunodeficiency virus (HIV)-discordant relationships. Papanicolaou smears were obtained in a prospective cohort of Kenyan HIV-discordant couples. Of 441 women, 283 (64%) were HIV-infected and 158 (36%) were HIV-uninfected with HIV-infected partners. Overall, 79 (18%) had low-grade and 25 (6%) high-grade cervical abnormalities. Male herpes simplex virus type 2 (HSV-2) seropositivity and lower couple socioeconomic status were associated with cervical abnormalities ( p 350 cells/µL) had the lowest prevalence of high-grade cervical lesions. HIV-infected women (CD4 > 350 cells/µL) and HIV-uninfected women with HIV-infected partners (CD4 ≤ 350 cells/µL) were at similar intermediate risk ( p > 0.05), and HIV-infected women (CD4 ≤ 350 cells/µL) had significantly higher risk of high-grade cervical abnormalities ( p = 0.05). Women in HIV-discordant relationships have high rates of cervical lesions and this may be influenced by couple-level factors, including HIV status and CD4 count of the infected partner.

Nutraceutically, much of the literature has indicated that an aglycon and its related glycoside would act similarly. However, controversial reports are accumulating. We hypothesize that rutin (RT) and quercetin (QT) pharmacodynamically could act differently. To confirm this, doxorubicin (DR) (8.5 mg/kg) was used to induce rat chronic kidney disease (CKD) and then treated with QT and RT (each 70 mg/kg body weight per day) for 13 weeks. QT exhibited better body weight gaining effect (420 ± 45) vs RT, 350 ± 57 g/rat (p0.001). DR raised the ratio kidney-to-body weight (%) to 0.82 (p0.001) vs RT, 0.62 (p0.01), and QT, 0.35 (p0.01). DR reduced the glomerular filtration rate to 25.2 vs RT, 48 ± 11.3; QT, 124.7 ± 12.8 (p0.001) and the control, 191.5 ± 15.7 mL/h (p0.001). DRCKD reduced hematocrit to 29 ± 5; RT, to 28 ± 5 (p0.05); QT, to 36 ± 6 vs the control 37.5 ± 4%, (p0.01). DRCKD reduced the serum albumin (s-Ab) to 2.1 ± 0.2 (p0.001); QT, to 2.7 ± 0.2 (p0.05) vs the normal 4.3 ± 0.5 g/dL, yet RT was totally ineffective. DRCKD raised serum cholesterol level to 340 ± 30; vs RT, 260 ± 12; QT, 220 ± 25; and the normal value, 70 ± 25 mg/dL. DRCKD increased serum triglyceride to 260 ± 15 (p0.001), RT and QT restored it to 170 ± 25 and 200 ± 15 (p0.05) vs the normal 26-145 mg/dL. DRCKD elevated blood urea nitrogen to 38 ± 3 vs RT, to 98 ± 6 mg/dL (p0.001), implicating "protein-energy malnutrition". RT stimulated serum creatinine (sCr) production to reach 6.0 ± 0.9 mg/dL (p0.001). QT did not alter the sCr level. RT but not QT induced uremia and hypercreatininemia. DR significantly downregulated Bcl-2, but highly upregulated Bax, Bad, and cleaved caspase-3, implicating the intrinsic mitochondrial pathway. DR damaged DNA, but QT completely rescued such an effect and recovered renal amyloidosis and collagen deposition. Conclusively, RT and QT act differently, and RT is inferior to QT with respect to treating CKD.

Human IgG2 antibodies contain three types of disulfide isoforms, classified by the number of Fab arms having disulfide links to the heavy chain hinge region. In the IgG2-B form, both Fab arms have interchain disulfide bonds to the hinge region, and in IgG2-A, neither Fab arm are disulfide linked to the hinge. The IgG2-A/B is a hybrid between these two forms, with only one Fab arm disulfide linked to the hinge. Changes in the relative levels of these forms over time are observed while IgG2 circulates in humans, suggesting IgG2-A→IgG2-A/B→IgG2-B conversion. Using a flow-through dialysis system, we studied the conversion kinetics of these forms in vitro under physiological conditions. For two IgG2κ antibodies, in vivo results closely matched the kinetics observed in vitro, indicating that the changes observed in vivo were solely conversions between isoforms, not differential clearance of specific forms. Moreover, the combined results validate the accuracy of the physiological model for the study of blood redox reactions. Further exploration of the conversion kinetics using material enriched in the IgG2-A forms revealed that the IgG2-A→IgG2-A/B rate was similar between IgG2κ and IgG2λ antibodies. In IgG2κ antibodies, conversion of IgG2-A/B→IgG2-B was slower than the IgG2-A→IgG2-A/B reaction. However, in IgG2λ antibodies, little IgG2-A/B→IgG2-B conversion was detected under physiological conditions. Thus, small differences in the C-terminus of the light chain sequences affect the disulfide conversion kinetics and impact the IgG2 disulfide isoforms produced in vivo.

We hypothesized that doxorubicin (DR) induced chronic kidney disease (CKD) could trigger the intrinsic and the extrinsic renal cell apoptotic pathways, while treadmill exercise could help prevent adverse effects. Male Sprague-Dawley rats were subjected to treadmill running exercise at a speed of 30 m/min, 30 or 60 min/day, 3 times per week, for a total period of 11 weeks. The physiological and biochemical parameters were seen substantially improved (DR-CKD control, 30 min, 60 min exercise): the ratio of kidney weight/body weight (0.89, 0.74, and 0.72); the WBC (1.35, 1.08, and 1.42 × 104 cells/μL); RBC (5.30, 6.38, and 6.26 × 106 cells/μL); the platelet count (15.1, 12.8, and 11.3 × 105/μL); serum cholesterol (659, 360, and 75 mg/dL); serum triglyceride (542, 263, and 211 mg/dL); BUN (37, 25, and 22 mg/dL). Bcl-2 and intramitochondrial cytochromecwere upregulated, while the levels of Bax, SOD, MDA, cleaved caspases 9, 3, 8, 12, and calpain were all downregulated in DRCKD groups with exercise. CHOP (GADD153) and GRP78 were totally unaffected. FAS (CD95) was only slightly suppressed in the 60 min exercise DRCKD group. Conclusively, exercise can ameliorate CKD through the regulation of the intrinsic and extrinsic apoptosis pathways. The 60 min exercise yields more beneficial effect than the 30 min counterpart.

Benign prostatic hyperplasia (BPH), an imbalance between androgen/estrogen, overexpression of stromal, and epithelial growth factors associated with chronic inflammation, has become an atypical direct cause of mortality of aged male diseases.Ginkgopossesses anti-inflammatory, blood flow-enhancing, and free radical scavenging effects. Considering strenuous exercise can reduce BPH risks, we hypothesizeGinkgo+ exercise (Ginkgo+ Ex) could be beneficial to BPH. To verify this, rat BPH model was induced by s.c. 3.5 mg testosterone (T) and 0.1 mg estradiol (E2) per head per day successively for 8 weeks, using mineral oil as placebo. Cerenin®8.33 μL/100 g was applied s.c. from the 10th to the 13th week, and simultaneously, Ex was applied (30 m/min, 3 times/week). In BPH,Ginkgoalone had no effect on T, 5α-reductase, and dihydrotestosterone (DHT), but suppressed androgen receptor (AR), aromatase, E2 and estrogen receptor (ER), and the proliferating cell nuclear antigen (PCNA); Ex alone significantly reduced T, aromatase, E2, ER, AR, and PCNA, but highly raised DHT. WhileGinkgo+ Ex androgenically downregulated T, aromatase, E2, and ER, but upregulated DHT, AR, and PCNA, implyingGinkgo+ Ex tended to worsen BPH. Conclusively,Ginkgoor Ex alone may be more beneficial thanGinkgo+ Ex for treatment of BPH.

The hypothalamic suprachiasmatic nucleus (SCN), a circadian pacemaker, is present in all mammalian brains. It has a complex organization of peptide-containing neurons that is similar among species, but calcium-binding proteins are expressed variably. Neurons containing calretinin have been described in the SCN in a number of species but not with association to circadian function. The objective of the present study is to characterize a calretinin neuron (CAR) group in the rat anterior hypothalamus anatomically and functionally with a detailed description of its location and a quantitative analysis of neuronal calretinin immunoreactivity at 3 times of day, 0600, 1400, and 1900 h, from animals in either light-dark or constant dark conditions. CAR neurons occupy a region in the dorsal and lateral SCN with a circadian rhythm in CAR immunoreactivity with a peak at 0600 h and a rhythm in cytoplasmic CAR distribution with a peak at 1400 h. CAR neurons should be viewed as an anatomical and functional component of the rat SCN that expands the definition from observations with cell stains. CAR neurons are likely to modulate temporal regulation of calcium in synaptic transmission.

Alzheimer’s disease is characterized by redistribution of the tau protein pool from soluble to aggregated states. Aggregation forms proteolytically stable core polymers restricted to the repeat domain, and this binding interaction has prion-like properties. We have compared the binding properties of tau and tubulin in vitro using a system in which we can measure binding affinities for proteins alternated between solid and aqueous phases. The study reveals that a phase-shifted repeat domain fragment from the Paired Helical Filament core contains all that is required for high affinity tau-tau binding. Unlike tau-tubulin binding, tau-tau binding shows concentration-dependent enhancement in both phase directions due to an avidity effect which permits one molecule to bind to many as the concentration in the opposite phase increases. Phosphorylation of tau inhibits tau-tau binding and tau-tubulin binding to equivalent extents. Tau-tau binding is favoured over tau-tubulin binding by factors in the range 19–41-fold, irrespective of phosphorylation status. A critical requirement for tau to become aggregation-competent is prior binding to a solid-phase substrate, which induces a conformational change in the repeat domain permitting high-affinity binding to occur even if tau is phosphorylated. The endogenous species enabling this nucleation event to occur in vivo remains to be identified. The findings of the study suggest that development of disease-modifying drugs for tauopathies should not target phosphorylation, but rather should target inhibitors of tau-tau binding or inhibitors of the binding interaction with as yet unidentified endogenous polyanionic substrates required to nucleate tau assembly.

Nanopores are powerful single-molecule sensors with nanometer scale dimensions suitable for detection, quantification, and characterization of nucleic acids and proteins. Beyond sequencing applications, both biological and solid-state nanopores hold great promise as tools for studying the biophysical properties of RNA. In this review, we highlight selected landmark nanopore studies with regards to RNA sequencing, microRNA detection, RNA/ligand interactions, and RNA structural/conformational analysis.

Chronic kidney disease (CKD) is usually associated with cardiac apoptosis and/or cardiac hypertrophy. We hypothesized that exercise can reduce the CKD-induced cardiac damage.The doxorubicin-induced CKD (DRCKD) model was used in rats to compare two exercise models: 60-min running and 60-min swimming. Results indicated that in healthy normal groups, the signals cardiotrophin-1 (CT-1), interleukin 6 (IL-6), leukaemia inhibitory factor receptor (LIFR), and gp130 were upregulated and janus kinase (JAK) and signal transducer and activation of transcription (STAT) were downregulated by both exercises. In contrast, all signals were highly upregulated in CKD. After exercise training, all signals (CT-1, IL-6, LIFR, gp130, and STAT) were downregulated, with JAK being only slightly upregulated in the running group but not in the swimming group. The myocyte death pathway (CT-1/IL-6 → LIFR/gp130 → PI3K → Akt → Bad) was excluded due to no change found for Bad. Nitric oxide (NO; normal, 15.63 ± 0.86 µmol/l) was significantly suppressed in CKD rats (2.95 ± 0.32 µmol/l), and both running and swimming training highly upregulated the NO level to 30.33 ± 1.03 µmol/l and 27.82 ± 2.47 µmol/l in normal subjects and 24.0 ± 3.2 µmol/l and 22.69 ± 3.79 µmol/l in the DRCKD rats, respectively. The endothelial progenic cells CD34 were significantly suppressed in DRCKD rats, which were not rescued significantly by exercise. In contrast, the CD 34 cells were only slightly suppressed in the healthy subjects by exercise.Both exercise regimens were beneficial by rescuing cardiac function in CKD victims. Its action mechanism was by way of inhibiting myocyte death and rescuing cardiac hypertrophy.